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Method and culture medium for enhanced detection of mycobacterium

A technology of mycobacteria and culture medium, applied in biochemical equipment and methods, microbial determination/inspection, bacteria, etc., can solve the problems of slow growth rate, hindering the detection of mycobacteria, etc.

Active Publication Date: 2012-05-23
BIO MERIEUX INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] However, detection of mycobacteria from clinical samples (e.g., sputum, lung fluid, tissue, or excreta) still represents an important biological challenge
One factor in this challenge arises from the fact that more rapidly growing bacteria may overgrow to suppress the slower growing mycobacterial organisms of interest, thus preventing or significantly hindering mycobacterial detection

Method used

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  • Method and culture medium for enhanced detection of mycobacterium
  • Method and culture medium for enhanced detection of mycobacterium
  • Method and culture medium for enhanced detection of mycobacterium

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0084] Example 1. TTD of different mycobacterial strains with and without fatty acid supplementation

[0085] To assess the effect of long-chain fatty acids (FA) on mycobacterial growth, fatty acid-free (FAF) BSA from Proliant, Inc. (Ames, Iowa) was selected. Based on fatty acid profile and growth performance, five long-chain fatty acids: myristic acid (C14:0), palmitic acid (C16:0), stearic acid (C18:0), oleic acid (C18:1) and linoleic acid (C18:2) was identified as a possible fatty acid supplement. Novel supplement formulations with FAF BSA were prepared and tested with and without 5 fatty acids (see Table 1 for novel supplement formulations). Target levels of fatty acid supplementation were based on fatty acid content obtained from previous assessments. The concentration of FAF BSA used was 10 g / L. Loss of fatty acids during filtration of the novel supplement was determined by fatty acid methyl ester (FAME) analysis. The recovery of fatty acids was observed to be >80% a...

Embodiment 2

[0097] Example 2. TTD of mycobacterial strains with and without α-ketoglutarate

[0098] To further improve the TTD of mycobacteria, the CO-producing pathway in mycobacterial cells was selected 2 Substrates and / or cofactors of enzymes including α-ketoglutarate, isocitrate, L-malate, oxaloacetic acid, lactate ) and L-arginine. Novel supplements with fatty acids were prepared and filter sterilized as described above. The substrates were added to the novel supplements at different concentrations.

[0099] Inspection 0.5×10 3 4 species of mycobacteria in CFU / ml. The effect of different concentrations of α-ketoglutarate on the growth (ie TTD of growth) of M. avium, M. intracellulare and M. tuberculosis cultures was studied. Using the new autoclavable BacT / ALERT MP flasks (as described above). 0.5 ml of the novel supplement with varying amounts of α-ketoglutarate was added to the novel BacT / ALERT before inoculating the MP flask with the mycobacterial culture MP culture flas...

Embodiment 3

[0104] Example 3. TTD of Mycobacterium species with conventional MAS

[0105] Through studies including polymyxin B (POLY B), amphotericin B (AMP B), nalidixic acid (NA), trimethoprim (TMP), azlocillin (AZL) and vancomycin (VAN ) of the 6 drugs determined the effectiveness of conventional MAS against mycobacterial growth. These studies were also performed to identify drugs and their concentrations that have adverse effects on mycobacterial growth.

[0106] For growth performance, conventional / old RFs were prepared and 6 antimicrobial agents were added at different concentrations. The selected concentration is a level 25-50% lower or higher than the concentration of conventional MAS, which is polymyxin B (POLY B) at 1000 units / ml, amphotericin B at 180 μg / ml ( AMP B), nalidixic acid (NA) at 400 μg / ml, trimethoprim (TMP) at 10.5 μg / ml, azlocillin (AZL) at 34 μg / ml, and vancomycin (VAN) at 5 μg / ml .

[0107] Growth of M. intracellulare, M. kansasii, M. scrofula, and M. tuberc...

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Abstract

The present invention relates to an improved culture medium and method for the enhanced growth and detection of Mycobacterium growth. The invention further relates to an improved mycobacterial reagent system or kit that can be used for the enhanced growth and detection of Mycobacterium.

Description

[0001] Cross References to Related Applications [0002] This application claims U.S. Provisional Patent Application No. 61 / 269,977, entitled "Method and Culture Medium for Enhanced Detection of Mycobacterium," filed July 1, 2009 No., which is incorporated into this article. field of invention [0003] The present invention generally relates to media and methods for enhanced detection of mycobacteria. More specifically, the present invention relates to various improvements in media and methods for improving or reducing the time to detection (TTD) of mycobacterial growth in media. Background of the invention [0004] Mycobacteria are a genus of bacteria characterized by acid-fast, non-motile, Gram-positive bacilli. The genus includes many species, including: Mycobacterium africanum, Mycobacterium avium (M.avium), Mycobacterium bovis (M.bovis), Mycobacterium bovis-BCG (M.bovis-BCG), M. chelonae, M. fortuitum, M. gordonae, M. intracellulare, M. kansasii ), Mycobacterium lep...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N1/38C12Q1/04
CPCG01N2333/35C12N1/38C12Q1/04C12N1/20C12Q1/045
Inventor 帕拉姆帕尔·多尔莱蒂西亚·巴顿约安尼·波尔蒂拉道格拉斯·劳沃恩
Owner BIO MERIEUX INC
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